In-cell architecture of an actively transcribing-translating expressome

Francis J O'Reilly, Liang Xue, Andrea Graziadei, Ludwig Sinn, Swantje Lenz, Dimitry Tegunov, Cedric Blötz, Neil Singh, Wim J. H. Hagen, Patrick Cramer, Jörg Stülke, Julia Mahamid, Juri Rappsilber

Research output: Contribution to journalArticlepeer-review


Structural biology performed inside cells can capture molecular machines in action within their native context. Here we developed an integrative in-cell structural approach using thegenome-reduced human pathogenMycoplasma pneumoniae. We combined whole-cellcrosslinking mass spectrometry, cellular cryo-electron tomography, and integrative modelingto determine an in-cell architecture of a transcribing and translating expressome at sub-nanometer resolution. The expressome comprises RNA polymerase (RNAP), the ribosome,and the transcription elongation factors NusG and NusA. We pinpointed NusA at theinterface between a NusG-bound elongating RNAP and the ribosome, and propose it canmediate transcription-translation coupling. Translation inhibition dissociatedthe expressome,whereas transcription inhibition stalled and rearranged it. Thus, the active expressomearchitecture requires both translation and transcription elongation within the cell.
Original languageEnglish
Pages (from-to)554-557
Number of pages4
Issue number6503
Publication statusPublished - 31 Jul 2020


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